Insertions in the protease (PR) region of human immunodeficiency virus (HIV) represent an interesting mechanism of antiviral resistance against HIV PR inhibitors (PIs). Here, we demonstrate the improved ability of a phosphonate-containing experimental HIV PI, GS-8374, relative to that of other PIs, to effectively inhibit patient-derived recombinant HIV strains bearing PR insertions and numerous other mutations. We correlate enzyme inhibition with the catalytic activities of corresponding recombinant PRs in vitro and provide a biochemical and structural analysis of the PR-inhibitor complex.

• MeSH
• HIV infekce farmakoterapie   virologie   MeSH
• HIV-1 chemie   účinky léků   enzymologie   genetika   MeSH
• HIV-proteasa chemie   genetika   metabolismus   MeSH
• inhibitory HIV-proteasy chemie   farmakologie   MeSH
• inzerční mutageneze * MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• molekulární modely MeSH
• organofosfonáty analýza   MeSH
• sekvence aminokyselin MeSH
• vazebná místa MeSH
• virová léková rezistence MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

While the selection of amino acid insertions in human immunodeficiency virus (HIV) reverse transcriptase (RT) is a known mechanism of resistance against RT inhibitors, very few reports on the selection of insertions in the protease (PR) coding region have been published. It is still unclear whether these insertions impact protease inhibitor (PI) resistance and/or viral replication capacity. We show that the prevalence of insertions, especially between amino acids 30 to 41 of HIV type 1 (HIV-1) PR, has increased in recent years. We identified amino acid insertions at positions 33 and 35 of the PR of HIV-1-infected patients who had undergone prolonged treatment with PIs, and we characterized the contribution of these insertions to viral resistance. We prepared the corresponding mutated, recombinant PR variants with or without insertions at positions 33 and 35 and characterized them in terms of enzyme kinetics and crystal structures. We also engineered the corresponding recombinant viruses and analyzed the PR susceptibility and replication capacity by recombinant virus assay. Both in vitro methods confirmed that the amino acid insertions at positions 33 and 35 contribute to the viral resistance to most of the tested PIs. The structural analysis revealed local structural rearrangements in the flap region and in the substrate binding pockets. The enlargement of the PR substrate binding site together with impaired flap dynamics could account for the weaker inhibitor binding by the insertion mutants. Amino acid insertions in the vicinity of the binding cleft therefore represent a novel mechanism of HIV resistance development.

• MeSH
• buněčné linie MeSH
• chemické modely MeSH
• difrakce rentgenového záření MeSH
• financování organizované MeSH
• HIV-1 enzymologie   fyziologie   genetika   MeSH
• HIV-proteasa genetika   chemie   izolace a purifikace   metabolismus   MeSH
• inhibitory reverzní transkriptasy chemie   MeSH
• inzerční mutageneze MeSH
• katalýza MeSH
• kinetika MeSH
• konsenzuální sekvence MeSH
• látky proti HIV terapeutické užití   MeSH
• ledviny cytologie   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• rekombinantní proteiny chemie   izolace a purifikace   metabolismus   MeSH
• replikace viru MeSH
• RNA virová analýza   MeSH
• sekvence aminokyselin MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• virová léková rezistence MeSH
• Check Tag
• lidé MeSH